Impact Goal: learn how to grow algae really well using a bioreactor that is very low maintenance
Secondary Goals: use our abundant algae for other projects like extracting oil, water purification, and a food source
11/4/16- We have been researching a lot about how to grow algae and how to build a bioreactor. So far, we plan on starting up our algae in a water jug with an aerating system and then once we figure out how it grows best, we'll move it to a bioreactor and see if it grows faster and more independently. Right now, we are working on getting the algae ordered. We are ordering chlorella algae, a species that is very tolerant of different growing conditions and wastewater. Our goal for the next few weeks is assembling a tank out of a water jug or container for the algae and getting the aerating system working. We also want to talk to students who have grown algae in the past and see what they say about how their algae grew.
Tubular Bioreactor: These bioreactors are designed to speed up the enrichment and growth of biomass such as algae. These long, clear tubes are pumped with dissolved CO2 to speed the rate of photosynthesis done by the algae. The algae produce lots more energy and become rich with oils (bio diesel) that can then be extracted by using a conventional oil press.
11/30/16:
This is a picture of our current system. We have two mason jars full of DI water covered with plastic wrap and tin foil. We have two incandescent bulbs to keep the system warm and an aerator to supply carbon dioxide. So far these have been our measurements:
Right now we are trying to figure out how we want to fertilize the algae. We have tried two different fertilizers, miracle grow and fertilizer ravioli (shultz boost packs). After testing the two fertilizers in different jars, there was a weird organism growing in the bottom of the jars.
12/9/16
We had more green stuff growing in the bottom of our jars, so we dumped them out and we are going to sterilize them in a glassware dishwasher somewhere in the school. Our algae is being ordered and hopefully it will be here early next week. We also got air stones to put at the end of our air lines to make the bubbles smaller and less violent.
12/14/16
Our algae is coming in a day or two. We have our jars in the dishwasher getting disinfected and we have enough DI water to fill them. We are also starting to gather supplies for the bioreactor. It looks like we are just going to have to find/buy a lot of tubing, but everything else we are finding ok. Here is what we have gathered so far:
and here is our current design for the bioreactor:
12/16/16
Our algae came yesterday! Dr. Tupper split it into two jars of fresh DI water. Today, we will try to get them to the appropriate temperature with our lights and also the appropriate nitrate levels with our Miracle Grow. We added 2.5 mL of Miracle Grow into each jar in hopes of reaching 40 ppm., maybe even a little more just for the weekend. Here it is: 12/19/16
Our algae survived the weekend and it even seems darker than it was. We are going to have to dilute it before break.
12/23/16
Our algae looked lighter and cloudier when we came in today. We think they may have run out of nutrients because we had a free yesterday and they only got a one day's dosage of food instead of two. We switched them into larger jars for the holidays and filled them with lots of food so they can hopefully survive the long week. Dr. Tupper says she will try to come in and feed them, so hopefully that will help. Here is a picture of the new big jars:
1/9/17-
We poured out our jars because the algae died. We should be getting more in this week. We have been working on building the bioreactor, and so far we have found some materials in the shop like tubing and connectors but we still need tubing for the algae part of the bioreactor and a tub for the lake. We have been getting ready to get the new algae, so we printed out a chart to record data on by the jars. We want to start keeping better track of what is being added to the jars and how that affects the jars.
1/12/17
New Algae arrived today. 20 mL water was removed from each mason jar, then 15 mL of the algae solution and 5 mL of miracle grow was added to each jar. The lights came on while the algae was being added. It looks like this:
1/13/17
We added 5mL miracle grow to the algae. Hopefully it will survive the long weekend. We have continued construction on our bioreactor, and once we get more parts next week we will be able to make serious progress on it. In the meantime, we have been looking at various ways to extract oil from algae. The cyclohexane and oil press strategies are very appealing. Fun Fact: algae has more oil per unit than any other plant.
1/19/17
Our algae is dying yet again. There are some major problems in our system that we must figure out. We will be doing some major research in these next few days to try to decide what changes need to be made so this stops happening. To begin with, we are ordering algal growth medium to get our algae started until we are able to move it into our own. Secondly, I think we have been adding way to much miracle grow. In years past, groups have only been adding 6 drops per liter while we having been adding 2.5mL to less than a liter. We are also going to have to invest in more sterilization techniques to sterilize our jars beforehand, because people have said that the milky white color we are observing is an infection of sorts. We will be trying to find out how to avoid this. In the meantime, we moved one of our jars into a larger jar in hopes of diluting the miracle grow a little bit. More research is to come. It is time to go back to the drawing board. This is a really helpful link: http://www.fao.org/docrep/003/w3732e/w3732e06.htm
1/27/17
Our algae is growing great! This week, we started one jar with bristol and metals growth medium, and one with DI water and four drops of miracle grow. It is now Friday and they are both very green and their nutrient levels are still looking strong. The problem was definitely overfeeding. Here is what our algae looks like now: (Jar A is DI water and four drops of miracle grow, and jar B is the bristol and metals growth medium).
1/30/17- Our algae is looking very good today. It got more green and dense over the weekend. We measured a higher level of nitrites in jar B (3 ppm) so we are trying to figure out where that came from and if that is going to be an issue.
2/3/17
This week our algae grew alot. The growth medium jar was very dark green, and our DI water jar only slightly less, so we decided to dilute. We split our two jars into four jars. The second DI water jar wasn't doing so well, so today we split the growth medium jar in half again to replenish the second DI jar. We also located a flourescent light and added it to our array. We covered the enclosure in more tin foil to prevent loss of light.
2/6/17
We ran into some problems over the weekend. Two jars died while two jars thrived. We're not quite sure why, but the two dead jars had significantly less nitrates than the thriving ones, which is weird because they were equal on Friday and were fed the same amount of food. Today we dumped the two dead jars and then split the other two up to have four jars again. They all look great! WE fed them 4 drops of MG each and hopefully they will all be good on Wednesday.
3/1/17
Our algae looked so dark green before break. We had six jars of varying sizes and they were all the darkest green we have ever seen. Unfortunately, our algae died over break. This could've been due to a loss of electricity or heat, or just the jars reaching their algae limit. We are going to start looking at the other aspects of our project in more detail until we get more algae. Perhaps we will look into more simpler bioreactors that use plastic bottles.
3/15/17
Last week we started putting together the new bioreactor plans . We assembled metal shelving and got a bunch of poland springs plastic bottles. Our algae is doing okay. We have two living small jars and one big jar and one of the small jars died over the snow day.
3/17/17
Today we got our bottle bioreactor going. Half our bottles have DI water and miracle grow and the other half have growth medium, and they are all inoculated at different densities. Unfortunately, someone turned off our power strip and some of our algae jars died. Here is our bottle bioreactor:
3/24/17
BUSINESS IS BOOMING!!!!!!!
3/27/17
We decided that the way we are going to extract algae is by french pressing it, microwaving it the break down the cell walls, then french pressing it again. Hopefully that will produce enough oil
3/31/17 Our extraction plans failed horribly when neither the french press filter nor a normal coffee filter could catch any significant amount of algae. As you can see in the picture, we cut the bottom off of gallons of milk and then stuck a coffee filter in and opened the lid so that we could pour our algae in and hopefully water would trickle out. It took a long time and a lot of algae just went straight through while any that did get caught basically became a part of the filter and was impossible to remove. So back to the drawing board on that. Otherwise, our algae is doing pretty well. Our jars are looking great. Our bottles, however, are having difficulties with the air supply, which puts a damper on algae growing. We will try to fix the problem next week by using air stones in our bottles too.
4/7/17
We added airstones to the air delivery system of our bottle bioreactor and they're working so well!!!! They are regulating the airflow so that all the bottle are receiving relatively the same amount of air and nobody is getting cut short. We filled all the bottles using algae from two dark mason jars so that those don't become over populated. Everything is fed (3 drops MG in each 16oz bottle) and we are expecting good things on Monday. Here is our current bottle setup:
5/11/17:
Term 4 Plan:
brian- explore/compare harvesting techniques and choose one suitable for greengineering
ariel- explore/compare harvesting techniques and choose one suitable for greengineering
anna- spirulina stuff
5/19/17-
Our algae is not doing too well right now. A few jars have either significantly lightened or died. The bottles in the bioreactor have lightened. We are still trying to figure out what might be causing our algae to decline. Besides from taking care of the algae, Brian has been working on the centrifuge that our team found. There seems to be a part that won't come unscrewed. Ariel has been researching lipid extraction methods. Anna is in the process of researching algae as a food source.
More on algae for food:
So far, I've found that Spirulina algae is the best algae to grow and eat because it has a larger shape and can be harvested more easily than chlorella. Spirulina can be bought online at places like carolina biological. There is a website called Spirulina Academy: http://www.spirulinaacademy.com/
And that site has a lot of good info on growing algae to eat. If we were to grow our own spirulina, we would need a tank or basin, a source of light (either sunlight or artificial light), a pump system, and a medium. For the medium, the site recommends the Zarrouk medium and dechlorinated tap water. The zarrouk medium is something we could make from the chemicals in the chem stock room, but it involves more than twenty different substances, so it would take a long time to mix. We could buy it online, but that is not a very sustainable option. Spirulina grows in 4-5 weeks and can be harvested with a fine filter. The fresh algae is safe to eat.
Lipid extraction: current research is being done on degrading the algae cell wall with the use of particular enzymes such as lysozyme, chitinase, and laminarase. Using these enzymes will degrade the cell wall well enough to increase lipid yields and extraction efficiency by a substantial margin. For the purposes of osmotic shock, which relies on the permeability of the cell wall, degrading the cell wall will expose the thin membrane and subsequently increase the efficiency of the process as a whole. In addition, killing the algae cells will permanently inhibit cell wall growth and repair, which will in turn help the degradation process and thus indirectly aid the extraction process as a whole. Currently, we are working on obtaining these enzymes and developing a concrete process or experiment to test the effectiveness of the process using osmotic shock.
5/24/17-
Today we dumped out a few dead jars and reorganized the live jars on our shelf. The bottles in the bioreactor are not looking too good. Brian is going to test the centrifuge using some of the dead algae jars. With so few classes left in the year, buying new algae is not a good option. We are just going to keep working with the algae we have left.
5/25/17-
Here is the centrifuge:
6/2/17
Last class we put a small amount of alum (<.1g) into a concentrated container of algae (~.8L). Within 20 mins, all the algae had dropped to the bottom!
We then poured off as much water as we could, and poured out the algae onto an aluminum foil tray and set it under a hot lamp. Today, the algae was fully dried, but it stuck to the foil a little so it was difficult to remove. We did remove a little tho and completed the harvesting process!
algae before:
Info from last year's team is here. Other years: 2012 - 2013, 2013 - 2014 and this very useful page, 2014 - 2015, and also here (from '14 - '15) 2015 - 2016
Impact Goal: learn how to grow algae really well using a bioreactor that is very low maintenance
Secondary Goals: use our abundant algae for other projects like extracting oil, water purification, and a food source
11/4/16- We have been researching a lot about how to grow algae and how to build a bioreactor. So far, we plan on starting up our algae in a water jug with an aerating system and then once we figure out how it grows best, we'll move it to a bioreactor and see if it grows faster and more independently. Right now, we are working on getting the algae ordered. We are ordering chlorella algae, a species that is very tolerant of different growing conditions and wastewater. Our goal for the next few weeks is assembling a tank out of a water jug or container for the algae and getting the aerating system working. We also want to talk to students who have grown algae in the past and see what they say about how their algae grew.
Tubular Bioreactor: These bioreactors are designed to speed up the enrichment and growth of biomass such as algae. These long, clear tubes are pumped with dissolved CO2 to speed the rate of photosynthesis done by the algae. The algae produce lots more energy and become rich with oils (bio diesel) that can then be extracted by using a conventional oil press.
11/30/16:
This is a picture of our current system. We have two mason jars full of DI water covered with plastic wrap and tin foil. We have two incandescent bulbs to keep the system warm and an aerator to supply carbon dioxide. So far these have been our measurements:
Right now we are trying to figure out how we want to fertilize the algae. We have tried two different fertilizers, miracle grow and fertilizer ravioli (shultz boost packs). After testing the two fertilizers in different jars, there was a weird organism growing in the bottom of the jars.
12/9/16
We had more green stuff growing in the bottom of our jars, so we dumped them out and we are going to sterilize them in a glassware dishwasher somewhere in the school. Our algae is being ordered and hopefully it will be here early next week. We also got air stones to put at the end of our air lines to make the bubbles smaller and less violent.
12/14/16
Our algae is coming in a day or two. We have our jars in the dishwasher getting disinfected and we have enough DI water to fill them. We are also starting to gather supplies for the bioreactor. It looks like we are just going to have to find/buy a lot of tubing, but everything else we are finding ok. Here is what we have gathered so far:
and here is our current design for the bioreactor:
12/16/16
Our algae came yesterday! Dr. Tupper split it into two jars of fresh DI water. Today, we will try to get them to the appropriate temperature with our lights and also the appropriate nitrate levels with our Miracle Grow. We added 2.5 mL of Miracle Grow into each jar in hopes of reaching 40 ppm., maybe even a little more just for the weekend. Here it is:
12/19/16
Our algae survived the weekend and it even seems darker than it was. We are going to have to dilute it before break.
12/23/16
Our algae looked lighter and cloudier when we came in today. We think they may have run out of nutrients because we had a free yesterday and they only got a one day's dosage of food instead of two. We switched them into larger jars for the holidays and filled them with lots of food so they can hopefully survive the long week. Dr. Tupper says she will try to come in and feed them, so hopefully that will help. Here is a picture of the new big jars:
1/9/17-
We poured out our jars because the algae died. We should be getting more in this week. We have been working on building the bioreactor, and so far we have found some materials in the shop like tubing and connectors but we still need tubing for the algae part of the bioreactor and a tub for the lake. We have been getting ready to get the new algae, so we printed out a chart to record data on by the jars. We want to start keeping better track of what is being added to the jars and how that affects the jars.
1/12/17
New Algae arrived today. 20 mL water was removed from each mason jar, then 15 mL of the algae solution and 5 mL of miracle grow was added to each jar. The lights came on while the algae was being added. It looks like this:
1/13/17
We added 5mL miracle grow to the algae. Hopefully it will survive the long weekend. We have continued construction on our bioreactor, and once we get more parts next week we will be able to make serious progress on it. In the meantime, we have been looking at various ways to extract oil from algae. The cyclohexane and oil press strategies are very appealing. Fun Fact: algae has more oil per unit than any other plant.
1/19/17
Our algae is dying yet again. There are some major problems in our system that we must figure out. We will be doing some major research in these next few days to try to decide what changes need to be made so this stops happening. To begin with, we are ordering algal growth medium to get our algae started until we are able to move it into our own. Secondly, I think we have been adding way to much miracle grow. In years past, groups have only been adding 6 drops per liter while we having been adding 2.5mL to less than a liter. We are also going to have to invest in more sterilization techniques to sterilize our jars beforehand, because people have said that the milky white color we are observing is an infection of sorts. We will be trying to find out how to avoid this. In the meantime, we moved one of our jars into a larger jar in hopes of diluting the miracle grow a little bit. More research is to come. It is time to go back to the drawing board. This is a really helpful link:
http://www.fao.org/docrep/003/w3732e/w3732e06.htm
1/27/17
Our algae is growing great! This week, we started one jar with bristol and metals growth medium, and one with DI water and four drops of miracle grow. It is now Friday and they are both very green and their nutrient levels are still looking strong. The problem was definitely overfeeding. Here is what our algae looks like now: (Jar A is DI water and four drops of miracle grow, and jar B is the bristol and metals growth medium).
1/30/17- Our algae is looking very good today. It got more green and dense over the weekend. We measured a higher level of nitrites in jar B (3 ppm) so we are trying to figure out where that came from and if that is going to be an issue.
2/3/17
This week our algae grew alot. The growth medium jar was very dark green, and our DI water jar only slightly less, so we decided to dilute. We split our two jars into four jars. The second DI water jar wasn't doing so well, so today we split the growth medium jar in half again to replenish the second DI jar. We also located a flourescent light and added it to our array. We covered the enclosure in more tin foil to prevent loss of light.
2/6/17
We ran into some problems over the weekend. Two jars died while two jars thrived. We're not quite sure why, but the two dead jars had significantly less nitrates than the thriving ones, which is weird because they were equal on Friday and were fed the same amount of food. Today we dumped the two dead jars and then split the other two up to have four jars again. They all look great! WE fed them 4 drops of MG each and hopefully they will all be good on Wednesday.
3/1/17
Our algae looked so dark green before break. We had six jars of varying sizes and they were all the darkest green we have ever seen. Unfortunately, our algae died over break. This could've been due to a loss of electricity or heat, or just the jars reaching their algae limit. We are going to start looking at the other aspects of our project in more detail until we get more algae. Perhaps we will look into more simpler bioreactors that use plastic bottles.
3/2/17
We researched different ways that people make homemade bioreactors, and a basic plastic bottle system seems the most popular. This link gives a really good explanation of one design: http://www.instructables.com/id/An-Algae-Bioreactor-from-Recycled-Water-Bottles/?ALLSTEPS
3/15/17
Last week we started putting together the new bioreactor plans . We assembled metal shelving and got a bunch of poland springs plastic bottles. Our algae is doing okay. We have two living small jars and one big jar and one of the small jars died over the snow day.
3/17/17
Today we got our bottle bioreactor going. Half our bottles have DI water and miracle grow and the other half have growth medium, and they are all inoculated at different densities. Unfortunately, someone turned off our power strip and some of our algae jars died. Here is our bottle bioreactor:
3/24/17
BUSINESS IS BOOMING!!!!!!!
3/27/17
We decided that the way we are going to extract algae is by french pressing it, microwaving it the break down the cell walls, then french pressing it again. Hopefully that will produce enough oil
3/31/17
Our extraction plans failed horribly when neither the french press filter nor a normal coffee filter could catch any significant amount of algae. As you can see in the picture, we cut the bottom off of gallons of milk and then stuck a coffee filter in and opened the lid so that we could pour our algae in and hopefully water would trickle out. It took a long time and a lot of algae just went straight through while any that did get caught basically became a part of the filter and was impossible to remove. So back to the drawing board on that. Otherwise, our algae is doing pretty well. Our jars are looking great. Our bottles, however, are having difficulties with the air supply, which puts a damper on algae growing. We will try to fix the problem next week by using air stones in our bottles too.
4/7/17
We added airstones to the air delivery system of our bottle bioreactor and they're working so well!!!! They are regulating the airflow so that all the bottle are receiving relatively the same amount of air and nobody is getting cut short. We filled all the bottles using algae from two dark mason jars so that those don't become over populated. Everything is fed (3 drops MG in each 16oz bottle) and we are expecting good things on Monday. Here is our current bottle setup:
5/11/17:
Term 4 Plan:
brian- explore/compare harvesting techniques and choose one suitable for greengineering
ariel- explore/compare harvesting techniques and choose one suitable for greengineering
anna- spirulina stuff
5/19/17-
Our algae is not doing too well right now. A few jars have either significantly lightened or died. The bottles in the bioreactor have lightened. We are still trying to figure out what might be causing our algae to decline. Besides from taking care of the algae, Brian has been working on the centrifuge that our team found. There seems to be a part that won't come unscrewed. Ariel has been researching lipid extraction methods. Anna is in the process of researching algae as a food source.
More on algae for food:
So far, I've found that Spirulina algae is the best algae to grow and eat because it has a larger shape and can be harvested more easily than chlorella. Spirulina can be bought online at places like carolina biological. There is a website called Spirulina Academy:
http://www.spirulinaacademy.com/
And that site has a lot of good info on growing algae to eat. If we were to grow our own spirulina, we would need a tank or basin, a source of light (either sunlight or artificial light), a pump system, and a medium. For the medium, the site recommends the Zarrouk medium and dechlorinated tap water. The zarrouk medium is something we could make from the chemicals in the chem stock room, but it involves more than twenty different substances, so it would take a long time to mix. We could buy it online, but that is not a very sustainable option. Spirulina grows in 4-5 weeks and can be harvested with a fine filter. The fresh algae is safe to eat.
Lipid extraction: current research is being done on degrading the algae cell wall with the use of particular enzymes such as lysozyme, chitinase, and laminarase. Using these enzymes will degrade the cell wall well enough to increase lipid yields and extraction efficiency by a substantial margin. For the purposes of osmotic shock, which relies on the permeability of the cell wall, degrading the cell wall will expose the thin membrane and subsequently increase the efficiency of the process as a whole. In addition, killing the algae cells will permanently inhibit cell wall growth and repair, which will in turn help the degradation process and thus indirectly aid the extraction process as a whole. Currently, we are working on obtaining these enzymes and developing a concrete process or experiment to test the effectiveness of the process using osmotic shock.
5/24/17-
Today we dumped out a few dead jars and reorganized the live jars on our shelf. The bottles in the bioreactor are not looking too good. Brian is going to test the centrifuge using some of the dead algae jars. With so few classes left in the year, buying new algae is not a good option. We are just going to keep working with the algae we have left.
5/25/17-
Here is the centrifuge:
6/2/17
Last class we put a small amount of alum (<.1g) into a concentrated container of algae (~.8L). Within 20 mins, all the algae had dropped to the bottom!
We then poured off as much water as we could, and poured out the algae onto an aluminum foil tray and set it under a hot lamp. Today, the algae was fully dried, but it stuck to the foil a little so it was difficult to remove. We did remove a little tho and completed the harvesting process!
algae before: